Site-directed mutagenesis to study the role of specific amino acids in the ligand binding domain of PPARs.

Details

Serval ID
serval:BIB_8287454AF822
Type
Article: article from journal or magazin.
Collection
Publications
Title
Site-directed mutagenesis to study the role of specific amino acids in the ligand binding domain of PPARs.
Journal
Methods in molecular biology
Author(s)
Mitro N., Gilardi F., Giudici M., Godio C., Scotti E., Crestani M.
ISSN
1940-6029 (Electronic)
ISSN-L
1064-3745
Publication state
Published
Issued date
2013
Peer-reviewed
Oui
Volume
952
Pages
137-144
Language
english
Notes
Publication types: Journal Article ; Research Support, Non-U.S. Gov't
Publication Status: ppublish
Abstract
The role of certain amino acids in the interactions of ligands with their cognate nuclear receptors is usually achieved by the resolution of the crystal structure of the receptor complexed with the ligand. As a complementary functional approach, site-directed mutagenesis, a technique broadly used in molecular biology, allows the assessment of the role of a specific amino acid in determining the interaction with a specific ligand. This method makes it possible to evaluate several mutations of a key amino acid for ligand binding and to determine the relationship between protein structure and ligand interaction. Here, we describe an application of this technique to evaluate different point mutations on the transcriptional activity of peroxisome proliferator-activated receptor γ (PPARγ) in the absence or presence of chemically different ligands.
Keywords
Amino Acids/metabolism, DNA Primers/genetics, Deoxyribonucleases, Type II Site-Specific/metabolism, Escherichia coli/cytology, Escherichia coli/genetics, HEK293 Cells, Humans, Ligands, Mutagenesis, Site-Directed/methods, Mutation, PPAR gamma/chemistry, PPAR gamma/genetics, PPAR gamma/metabolism, Plasmids/genetics, Protein Structure, Tertiary, Transformation, Genetic
Pubmed
Create date
21/03/2019 11:29
Last modification date
20/02/2020 6:26
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